Dear there,
From the TCGAbiolinks paper (https://journals.plos.org/ploscompbiol/article?id=10.1371/journal.pcbi.1006701), I see it need scaling Recount2 data because it was sequenced using Rail-RNA. Here is the code they used as below for downstream differential expression analysis (DEA) using "TCGAanalyze_DEA". They also used "TCGAanalyze_Normalization" and "TCGAanalyze_Filtering" to do the normalization and filtering. Since DESeq2 takes the raw read counts as input, I am wondering do I need any of these 3 (scaling, normalization, and filtering) if I used "TCGAquery_recount2" to download GTEx and TCGA data? Thanks a lot!
Xiaofei
#####Preparing/scaling Recount2 data because it was sequenced using Rail-RNA
eset.gtex<-assays(scale_counts(ucs.recount.gtex$GTEX_uterus, round = TRUE))$counts
eset.tcga<-assays(scale_counts(ucs.recount.tcga$TCGA_uterus, round = TRUE))$counts